Establishment of B cell hybridomas with B cell surface antigens.
M12.4, one of several B lymphomas derived from BALB/c mice, was mutagenized with ethyl methanesulfonate in vitro. M12.4.1, a subline of the mutant cells, was sensitive to hypoxanthine-aminopterin-thymidine selective medium and was fused with normal splenic B lymphocytes of C57BL/6ByJ mice. The hybridomas obtained from this fusion were shown to express mu heavy chain, H-2KbDb, Iad, and Iab on the cell surface by analyses of flow microfluorometry and a cytotoxicity assay, although parental M12.4.1 lacked mu heavy chain on the cell membrane. These results demonstrate that the B cell hybridomas with B cell surface antigens have been established in vitro. IgM molecules on the cell surface of the hybridomas were shown to originate from normal B cells of C57BL/6ByJ mice by flow microfluorometry analyses after staining with fluorescein-labeled Bet 1, a monoclonal rat antibody that recognizes Igh-6.5, a mouse IgM allotypic determinant. These hybridomas could generate IgM-secreting cells at the high frequency (more than 10% of the cultured cells) when stimulated with bacterial lipopolysaccharide. On the other hand, parental M12.4.1 did not develop any IgM-secreting cells under the same conditions. These findings suggest that these B cell hybridomas with B cell surface antigens may be a good model for the study of B cell differentiation.
D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000276:immunology*; D006183:H-2 Antigens / Q000032:analysis; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D006825:Hybridomas / Q000276:immunology*; D007075:Immunoglobulin M / Q000096:biosynthesis; D007148:Immunoglobulin mu-Chains / Q000032:analysis; D008223:Lymphoma / Q000276:immunology; D051379:Mice; D013154:Spleen / Q000276:immunology
D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D006183:H-2 Antigens; D000949:Histocompatibility Antigens Class II; D007075:Immunoglobulin M; D007148:Immunoglobulin mu-Chains